Theoretical approach to the dynamics of the inoculum density of Verticillium dahliae in the soil: first test of a simple model

A mathematical equation was developed that describes the inoculum densities of microsclerotia of Verticillium dahliae in the soil over a long time span. The equation was based on measurable parameters and ecologically meaningful principles. In the model, the number of systemic infections of plant roots during crop growth was related to soil inoculum density. In turn, formation of microsclerotia in debris and reduction of the amount of crop growth were related to the number of systemic infections. Finally, a gradual release and mortality of microsclerotia in the soil were included to calculate subsequent inoculum densities in the soil.
Fitting the function to experimental data of potato cvs Element, Ostara, Mirka and Astarte, flax, pea, barley, sugar beet, onion and faba bean gave a very high correlation between observed and predicted soil inoculum densities. The clear differences in inoculum production among potato cultivars and other crops were expressed in quantitative terms. The highest inoculum density after incorporation of the debris of a susceptible crop was estimated to occur at 2.3 thermal time units of 3600 degree days (base 0°C). Ten per cent of the initial input of inoculum was still present after 4.5 thermal time units. The model was used to predict the dynamics of soil inoculum densities for V. dahliae under various cropping frequency schemes and performed satisfactorily.     

Relationships Between Verticillium dahliae Inoculum Density and Wilt Incidence, Severity, and Growth of Cauliflower

ABSTRACT Microplot and field experiments were conducted to evaluate the effects of inoculum density on Verticillium wilt and cauliflower growth. Soil containing Verticillium dahliae microsclerotia was mixed with various proportions of fumigated soil to establish different inoculum densities (fumigated soil was used as the noninfested control). Seven inoculum density treatments replicated four times were established, and the treatments were arranged in a randomized complete block design. Soil was collected from each microplot immediately after soil infestation for V. dahliae assay by plating onto sodium polypectate agar (NP-10) selective medium using the Anderson sampler technique. Five-week-old cauliflower was transplanted into two beds within each 1.2- by 1.2-m microplot. At the same time, several extra plants were also transplanted at the edge of each bed for destructive sampling to examine the disease onset (vascular discoloration) after planting. Cauliflower plants were monitored for Verticillium wilt development. Stomatal resistance in two visually healthy upper and two lower, diseased leaves in each microplot was measured three times at weekly intervals after initial wilt symptoms occurred. At maturity, all plants were uprooted, washed free of soil, and wilt incidence and severity, plant height, number of leaves, and dry weights of leaves and roots were determined. The higher the inoculum density, the earlier was disease onset. A density of 4 microsclerotia per g of dry soil caused 16% wilt incidence, but about 10 microsclerotia per g of soil caused 50% wilt incidence. Both wilt incidence and severity increased with increasing inoculum density up to about 20 microsclerotia per g of soil, and additional inoculum did not result in significantly higher disease incidence and severity. A negative exponential model described the disease relationships to inoculum levels under both microplot and field conditions. Stomatal resistance of diseased leaves was significantly higher at higher inoculum densities; in healthy leaves, however, no treatment differences occurred. The height, number of leaves, and dry weights of leaves and roots of plants in the fumigated control were significantly higher than in infested treatments, but the effects of inoculum density treatments were variable between years. Timing of cauliflower infection, crop physiological processes related to hydraulic conductance, and wilt intensity (incidence and severity) were thus affected by the inoculum density. Verticillium wilt management methods used in cauliflower should reduce inoculum density to less than four micro-sclerotia per g of soil to produce crops with the fewest number of infected plants.     

Detection of Colletotrichum coccodes and Helminthosporium solani in soils by bioassay

The sensitivity of a bioassay in detecting soil inoculum of Colletotrichum coccodes and Helminthosporium solani was examined using potato minitubers and microplants. Tests were conducted on soils which were collected from fields in which the interval after a previous potato crop differed, and which were also artificially infested with conidia or microsclerotia. For C. coccodes , determining plant infection based on the occurrence of infected roots after 9–12 weeks was a sensitive method for detecting and quantifying the amount of inoculum in soil. Infestations of less than 0·4 microsclerotia per g soil were detected in artificially infested soils. A semiselective medium, developed for isolating C. gloeosporioides from pepper, detected soil infestations by C. coccodes as low as nine conidia or one microsclerotium per g soil in artificially infested soil. For H. solani , infection on minitubers was a sensitive measure, with soil inoculum of fewer than 10 conidia per g soil being detected. Soil infestation could be quantified by assessing the percentage surface area of minitubers covered by sporulating lesions, which was strongly related to the amount of soil infestation. The results of these bioassay tests were compared with published results for real-time quantitative PCR assays on the same soils. The two methods were in good agreement in artificially infested soils, but the bioassay appeared to be more sensitive with naturally infested soils.     

The potential for the Rapid Screening of Potato Cultivars (Solanum tuberosum) for Resistance to Powdery Scab (Spongospora subterranea) using a Laboratory Bioassay

Powdery scab of potato, once established in a field, is difficult to control because of the longevity of the resting spores (cystosori) of the causal organism, Spongospora subterranea f.sp. subterranea. Host resistance is likely to be the most efficient in a long-term control strategy for preventing build-up of field inoculum and spread of the disease. Resistance screening of potato cultivars is mostly done in laborious field trials where disease development is likely to be unpredictable. A bioassay with potato tissue cultured plantlets and cystosori as inoculum is described and was tested for its potential to screen potato cultivars at an early stage for their relative susceptibility to powdery scab by comparing the lab results with field data. With cystosori inoculum of Swiss origin, the laboratory test showed clear differences between the potato cultivars in the severity of zoosporangial root infection which correlated better with ranked tuber infection data, compared to root galling. There are apparent differences in the relative trends in susceptibility between roots and tubers of five selected cultivars when using naturally infested soil instead of prepared cystosori as inoculum in the lab bioassay. Furthermore, differences in the severity of zoosporangial root infection of two selected cultivars were found when cystosori from different countries where used as inoculum. A possible host genotype × pathogen interaction is discussed. The bioassay has the potential to screen and select for resistant material at an early breeding stage thus making field trials not unnecessary but more economical. It will allow the use of a standard set of pathogen collections and facilitate testing for inoculum virulence in infested soils.     

Suppression of potato cyst nematode root penetration by the endoparasitic nematophagous fungusHirsutella rhossiliensis

The endoparasitic nematophagous fungusHirsutella rhossiliensis was tested for its ability to suppress root penetration and cyst formation by the potato cyst nematode speciesGlobodera pallida. Isolates ofH. rhossiliensis were obtained from infected potato cyst nematode juveniles from different starch potato fields in The Netherlands. The isolates showed no difference in spore adhesion to juveniles on agar plates (adhesion rate: ±90%). The most rapid growing isolate, CBS 108.94, was used for experiments. Vegetative mycelial colonies ofH. rhossiliensis CBS 108.94, grown in potato dextrose broth, were used as soil inoculum. During submerged cultivation the mycelial colonies produced phialides (spore-bearing cells) but no spores. Exposed to the air, however, spores were rapidly formed. The effect of different soil inoculum densities of mycelial colonies on root penetration byGlobodera pallida was examined in an experiment in 250-ml pots. Up to a mycelial colony concentration representing a potential spore density of 10⁴ g^–1 soil no suppression occurred. At approximated densities of 2.5×10⁴ and 10⁵ spores g^–1 soil the numbers of juveniles which penetrated roots were reduced by 30% and 34%, respectively. The distribution of the inoculum could be improved by fragmentation of the mycelial colonies before soil inoculation. Using mycelial fragments, again no suppression of root penetration was observed up to a potential spore density of 10⁴ g^–1 soil, but at densities of 10⁵ and 10⁶ g^–1 a suppression of 54% and 88%, respectively, was measured. In a greenhouse experiment, soil inoculation with mycelial colonies with a potential spore production of 2.5×10⁵ g^–1 soil resulted in a suppression of root penetration of 37% and 51% after 5 and 6 weeks, respectively, but the number of newly formed cysts after 18 weeks in soil was not different for control and inoculated pots. It is concluded thatH. rhossiliensis may be useful for the reduction of root damage caused by juveniles of potato cyst nematodes, but the usefulness for population control is doubtful.     

Fitness cost but no selection for virulence in Meloidogyne incognita after two consecutive crops of eggplant grafted onto Solanum torvum

The eggplant Solanum melongena cv. Cristal, either ungrafted or grafted onto the Solanum torvum rootstock cv. Brutus, was cultivated for two consecutive years in the same plots in a plastic greenhouse to assess the level of resistance to Meloidogyne incognita and crop yield. At the end of the second crop, the putative selection for virulence of the nematode subpopulations coming from infected ungrafted and grafted eggplant was assessed in the eggplant and in S. torvum in a pot experiment. Nematode population densities at transplantation in 2017 ranged from 2 to 378 per 100 cm³ of soil and did not differ between ungrafted and grafted eggplant. At the end of each crop, a higher galling index and number of nematodes in soil and in roots was registered in ungrafted compared to grafted eggplant. The grafted eggplant was categorized as resistant in 2017 and as highly resistant in 2018. Eggplant yield did not differ irrespective of grafting in 2017 after being cultivated for 135 days, but it differed after 251 days of cultivation in 2018. In the pot experiment, S. torvum was categorized as resistant to both M. incognita subpopulations. However, the M. incognita subpopulation obtained from roots of S. torvum produced 49.4% fewer egg masses and 56% fewer eggs per plant in the eggplant than the nematode subpopulation obtained from roots of the eggplant cv. Cristal. The results of this study revealed that the infective and reproductive fitness of the nematode decreased without having been selected for virulence.     

The influence of cyst nematodes and drought on potato growth

In two experiments in the Wageningen Rhizolab with potato cv. Mentor planted in soils with or without potato cyst nematodes (Globodera pallida, W) the number of roots per cm² was observed two weekly by video camera, in horizontally placed minirhizotrons at depths varying from 5 to 100 cm. In both experiments initial root growth was more rapid in the top soil of the uninfected soil. In the first experiment under optimal water supply, root formation continued longer in the top 30 cm of infested soil leading to twice as many roots at the end of the growing season as in uninfested soil. In the subsoil from 30 to 100 cm, however, root formation was strongly reduced by cyst nematodes leading to an uneven distribution of roots throughout the profile. In the second experiment potato cyst nematodes only increased rooting in the top soil with reduced irrigation. Potato cyst nematodes did not affect the water use efficiency of the crop whereas reduced irrigation increased water use efficiency by about 22%. Without potato cyst nematodes the soil profile was depleted of mineral nitrogen until a depth of 1 m whereas with high initial population densities no nitrogen was taken up in the subsoil between 30 and 100 cm. The spational heterogenity of roots and nitrogen in the soil is an important mechanism of damage. This finding may lead to improved cultural practices and breeding for tolerance.     

Effect of <Emphasis Type="Italic">Meloidogyne ethiopica</Emphasis> parasitism on water management and physiological stress in tomato

Root-knot nematodes (RKN) are obligate endoparasites that severely damage the host root system. Nutrient and water uptake are substantially reduced in infested plants, resulting into altered physiological processes and reduced plant growth. The effect of nematode infestation on the morphological changes of roots and subsequent physiological plant responses of infested tomatoes with the RKN Meloidogyne ethiopica was studied in a pot experiment. Plants were infested with two inoculum densities (10 or 50 eggs per cm³ substrate) and its effect was evaluated 74 and 102 days post inoculation (DPI). Morphological changes and root growth was determined by analysing scanned images of the whole root system. Nematode infestation reduced the portion of fine roots and increased that of coarse roots due to gall formation. Fine roots of non-infested control plants represented around 51% of the area of the whole root system at 74 and 102 DPI. In comparison to controls, plants inoculated with low and high nematode density had 2.1 and 3.2-times lower surface area of fine roots at 102 DPI. Root analyses revealed that plants had a very limited ability to mitigate the effects of the root-knot nematodes infestation by altering root growth. Root galls had a major influence on the hydraulic conductivity of the root system, which was significantly reduced. The low leaf water potential of infested plants coincided with decreased stomatal conductivity, transpiration and photosynthesis. The latter two were reduced by 60–70% when compared to non-infested control plants.     

Effects of crop rotation and removal of crop debris on the soil population of two isolates of Verticillium dahliae

Microsclerotia of Verticillium dahliae are produced in large numbers on senescing parts of host plants and remain viable in the soil for many years. Changes in the population density, i.e. density of microsclerotia, in the soil were measured in micro-plots using two isolates of V. dahtiae , specific to either field bean or potato, several crop sequences comprising potato, field beans and barley, and either the removal of aerial debris of the crops or incorporation into soil.
Potato was more susceptible to the potato isolate and field bean more susceptible to the field bean isolate. Removal of debris of potato and field bean reduced numbers of microsclerotia in the soil in the subsequent years, but removal of barley straw had no effect. Initially non-infested control micro-plots became infested, probably by the growth of potato roots into the naturally infested subsoil. The rate of increase of the microsclerotial population in the non-infested control micro-plots was larger than in the initially infested treatments, because more colonized debris was produced. It is concluded that removal of aerial debris of host crops is important to reduce the soil population of V. dahtiae.     

Infectivity,inoculum density and germination of Spongospora subterranea resting spores: a solution-culture test system1

Spongospora subterranea, causal agent of powdery scab of potatoes and vector of potato mop-top furovirus, survives in the soil as balls of resting spores (cystosori). So far, the factors affecting longevity, germination and infectivity of cystosori have not been investigated. A rapid and versatile bioassay with tomatoes as bait plants has been developed to quantify the infectivity of cystosorus inoculum or infested soil. The intensity of root infection, as a measure of infectivity, was determined by evaluating the quantity of zoosporangia present in epidermal cells and root hairs of the whole, stained root system. A correlation was obtained between the intensity of root infection and the cystosorus inoculum density in nutrient solution. Sterile soil suppressed the inoculum potential of pure cystosori. Infectivity of untreated soil decreased with increasing time of storage. Root infection was not influenced by the pH level of the nutrient solution.     

Interactions Between the Potato Cyst Nematode Globodera rostochiensis and Diseases Caused by Rhizoctonia solani AG3 in Potatoes Under Field Conditions

Findings from 2 years of field experiments investigating the relationship between Globodera rostochiensis and Rhizoctonia solani on unique field sites are reported. In 2000, a field experiment was positioned on land that had previously been used for experimental work investigating integrated potato cyst nematode (PCN) management methods. This study had produced an ‘untypical’ mosaic of PCN population densities ranging from 5 to 221 eggs g⁻¹ soil. In 2001, the field experiment was conducted on a different field site and overlaid on a focus of G. rostochiensis population densities ranging from 11 to 108 eggs g⁻¹ soil. In each experiment, potatoes (cv. Désirée) were grown in plots with similar population densities of G. rostochiensis that were either uninoculated or inoculated with R. solani. A series of potato plant harvests were undertaken to investigate the effects of nematode infestation on the incidence and severity of R. solani diseases and the associated development of plants. In both experiments, a clear relationship was found between the density of G. rostochiensis juveniles present in potato roots and the incidence of stolons infected by R. solani, 6 weeks after planting. For the first time this interaction has been determined under field conditions. The results of the study suggest that the interaction between nematode and fungus is indirect and possible mechanisms are discussed.     

Evaluation ofAmaranthus sp. andVernonia amygdalina, and soil amendments with poultry manure for the management of root-knot nematodes on eggplant

The suppressive effect of vernonia (Vernonia amygdalina), amaranth (Amarathus sp.) and poultry manure on root-knot nematodes (RKNs) (Meloidogyne spp.) infecting eggplant (Solanum macrocarpon) was studied at two sites in southern Benin naturally infested with these nematodes. After 3 months, soil and root-inhabiting RKN populations were significantly less (P≦0.05) in the plots cropped with vernonia, amaranth, and eggplant amended with poultry manure (PM) at the rate of 40 t ha⁻¹ as compared with the rate of 20 t ha⁻¹ and with the control. Poultry manure was more effective after 2 months than after 3 months. Overall, vernonia was the most effective treatment affecting RKN populations in the roots and the soil. The use of these treatments in nematode management through rotation and co-planted crops is discussed. http://www.phytoparasitica.org posting August 6, 2008.     

Mechanism of broccoli-mediated verticillium wilt reduction in cauliflower

ABSTRACT Broccoli is resistant to Verticillium dahliae infection and does not express wilt symptoms. Incorporation of broccoli residues reduces soil populations of V. dahliae. The effects of broccoli residue were tested on the colonization of roots by V. dahliae, plant growth response, and disease incidence of both broccoli and cauliflower in soils with different levels of V. dahliae inoculum and with or without fresh broccoli residue amendments. The three soils included a low-Verticillium soil, a high-Verticillium soil, and a broccoli-rotation soil (soil from a field after two broccoli crops) with an average of 13, 38, and below-detectable levels of microsclerotia per g of soil, respectively. Cauliflower plants in broccoli-amended high-Verticillium soil had significantly (P 相似文献   

甘薯对马铃薯腐烂茎线虫趋化性的影响

由马铃薯腐烂茎线虫侵染造成的甘薯茎线虫病是一种严重影响甘薯产量和品质的病害。线虫向寄主根部移动及取食危害可能是受寄主释放的化感信息物质控制。为最终明确甘薯中对线虫具有高效引诱能力的物质,本研究利用水琼脂平板法测试了甘薯不同品种与不同组织对线虫的引诱作用。结果表明不同品种甘薯薯块和薯茎对马铃薯腐烂茎线虫引诱能力具有一定差异,感线虫品种栗子香的薯块对线虫的引诱率要高于其他测试品种。随着薯块/薯茎在水琼脂平板上放置时间延长,其吸引线虫的效果增加,96 h后引诱率达到20%。同一品种甘薯茎的引诱能力显著高于薯块、薯叶。甘薯中所含对线虫具引诱作用的活性物质对热稳定,80℃加热8 h后薯茎对线虫引诱率与新鲜茎相比无显著差异,为4.5%。甘薯茎经过不同有机溶剂分别萃取后,乙醇萃取物对线虫的引诱率为6.7%,显著高于其他萃取物。本文初步明确了甘薯中所含线虫趋化物的性质,为其开发应用奠定了基础。     

Een virus inAtropa belladonna

Summary In a bgarden at Baarn, The Netherlands,Atropa belladonna plants, grown from seed, showed symptoms similar to those described bySmith (1946, 1957) for Belladonna mosaic. After inoculation of solanaceous test plants with sap from diseased plants, the following species showed symptoms:Atropa belladonna L.,Capsicum annuum L.,Hyoscyamus niger L.,Nicandra physaloides Gaertn.,Nicotiana glutinosa L.,Nicotiana tabacum L. var. Samsun,Petunia hybrida andPhysalis floridana Rydb. The symptoms suggest that the virus may be identical with that described bySmith. A high virus concentration was found inHyoscyamus niger. Nicandra physaloides, Petunia hybrida, Physalis floridana, in the roots and the pericarp of diseasedAtropa plants, and also inSolanum nigrum L. andDatura stramonium L. The latter two species showed hardly any symptoms. A. low virus concentration was found inCapsicum annuum, though this plant showed severe symptoms.The dilution end point of the virus was between 10^–3 and 10^–4; virusinactivation occurred between 70° and 80°C.In electron micrographs the rod-shaped virus particles appeared similar to those of rattle virus.Virus could be detected in the roots of tobacco plants after the leaves had been inoculated with sap of diseasedAtropa-plants (Table 1). The reverse did not occur. Following immersion of the roots of tobacco plants in virus-containing sap these plants were potted in steamed soil. Subsequently the roots proved to be infected but the stems and leaves contained no virus. However,Atropa plants treated in the same way, did show leaf symptoms.It appeared, that the roots of young, healthy tobacco plants could become infected with virus, when grown in naturally infested soil for only tow days (Table 2). Fungus cultures isolated from diseased roots did not show any infectivity. Nematodes are probably the vectors of this virus (Sol, Van Heuvel & Seinhorst, 1960).Met medewerking van Merr.J. M. Dekhuyzen-Maasland, Dr. S. Gayed (Karthoum), Mej.C. van Heuven, C. de Vooys en Mej.R. van Wessem.     

Manipulating inoculum densities of Verticillium dahliae and Pratylenchus penetrans with green manure amendments and solarization influence potato yield

We used cover crops with demonstrated efficacy against Verticillium dahliae and Pratylenchus penetrans in combination with the biocidal practice of solarization to determine the importance of targeting both organisms for managing potato early dying, an issue relevant to the search for alternatives to soil fumigation. Two experiments were conducted in commercial fields using a split-plot design with cover crop treatments of rapeseed, marigold, forage pearl millet, sorghum-sudangrass, and corn as the main plot factor and solarization as the subplot factor. Cover crops were grown and solarization applied in year one, followed by potato in year two. The main effect of solarization was significant for reduced inoculum levels of both organisms in year two and increased tuber yields. The main effect of cover crop was also significant with lower population densities of P. penetrans following the marigold and millet treatments and of V. dahliae following rape and sorghum-sudangrass. The cover crop treatments influenced yield in only one of the experiments in the absence of solarization. The combinatorial effect of cover crops and solarization resulted in a wide range of pathogen population densities. Mean soil inoculum levels were negatively related to yield for V. dahliae in experiment 1, and for P. penetrans and the P. penetrans × V. dahliae interaction in both experiments.     

大丽轮枝菌在茄田土壤中的分布及抽样方法

对茄子黄萎病在田间的空间分布及取样方法进行了研究,并探讨了田间土壤带菌量的准确测定方法。研究结果表明:用分布型指数法测定茄田土壤中微菌核的空间分布型,当菌核数量在8-63 ～39-93 个/g 干土范围内,为聚集分布。当茄田发病率小于75 % 时,病株在田间均属聚集分布;发病率大于75 % 时呈均匀分布。土壤中微菌核的调查方法以棋盘式为最适宜。     

大丽轮枝菌在茄田土壤中的分布及抽样方法

对茄子黄萎病在田间的空间分布及取样方法进行了研究,并探讨了田间土壤带菌量的准确测定方法。研究结果表明:用分布型指数法测定茄田土壤中微菌核的空间分布型,当菌核数量在8-63 ～39-93 个/g 干土范围内,为聚集分布。当茄田发病率小于75 % 时,病株在田间均属聚集分布;发病率大于75 % 时呈均匀分布。土壤中微菌核的调查方法以棋盘式为最适宜。     

Ralstonia solanacearum causing potato bacterial wilt: host range and cultivars’ susceptibility in Rwanda

Members of the Ralstonia solanacearum species complex (RSSC), causing potato bacterial wilt or brown rot, are highly contagious and there are no known cultivars with durable resistance to the pathogen. This study hypothesized (a) that crops intercropped or rotated with potato, plants in the same family, and plants grown in the neighbouring fields can host the pathogen and they can be potential sources of primary inoculum, and (b) that potato cultivars currently multiplied by the public tissue culture laboratory in Rwanda are less susceptible to the pathogen. Fourteen plant species and potato, and nine potato cultivars were tested for susceptibility to an RSSC phylotype II strain under greenhouse conditions. The bacteria induced symptoms on potato, tomato, tree tomato, sweet pepper, and eggplant only. Among the plant species with symptoms, potato, tomato, and tree tomato wilted completely. There was a significant difference in days to symptom expression and to complete wilting (p < .0001). While all tested potato cultivars were found to be susceptible, the number of days to first symptom expression, days to complete wilting, area under the disease progress curve (AUDPC), and the number and weight of harvested tubers varied considerably. Cultivars Cruza, Kinigi, and CIP-58 were less susceptible whereas the cultivars Gikungu, Kirundo, and Victoria were highly susceptible. There is a strong need to search for other sources of resistance. The results indicate that some plant species that might serve as a reservoir of the bacterium should be avoided in the vicinity of potato crops.     

Fusarium solani f.sp.cucurbitae andF. oxysporum f.sp.niveum inoculum densities in tunisian soils and their effect on watermelon seedlings

Populations ofFusarium solani f.sp.cucurbitae (Fsc) andFusarium oxysporum f.sp.niveum (Fon) in naturally infested soil of watermelon fields were counted by the soil dilution method with subsequent pathogenicity tests. Inoculum density varied within the same region from one field to another, ranging between 9 and 1600 CFU g^?1 soil forFsc and from 0 to 200 CFU g^?1 soil forFon. Fusarium crown- and root-rot-diseased seedlings were observed in most soils (93%); however, Fusarium wilt was observed in only 34% of soil samples. The disease incidence on cv. ‘Giza’ (Y) increased significantly with inoculum density in the soil (X) (P<0.001). ForFsc, the relationship between inoculum density and disease incidence was characterized by the equation Y=0.0005X+0.165 (R²=0.67). ForFon, the equation was Y=0.003X?0.0014 (R²=0.88). Based on these equations, the estimated inoculum densities required to cause 50% disease incidence (DI₅₀) on cv. Giza plants was 670 and 171 CFU g^?1 soil forFsc andFon, respectively.